Key features and details
- Platform: Microplate reader, Fluor. microscope, Flow cyt.
- Sample type: Adherent cells, Suspension cells
Product nameCellular ROS Assay Kit (Orange)
See all Oxidative Stress kits
Sample typeAdherent cells, Suspension cells
Cellular ROS Assay Kit (Orange) ab186028 uses an ROS sensor to quantify ROS in live cells. The orange dye used in the ROS assay protocol is cell-permeable and generates orange fluorescence when it reacts with ROS. The fluorescence signal of the dye can be measured by fluorescence microscopy, high-content imaging, microplate fluorometry, or flow cytometry.
ab186028 provides a sensitive fluorometric, one-step assay to detect intracellular ROS (especially superoxide and hydroxyl radical) in live cells within 1 hour. The ROS assay can be performed in a convenient 96-well or 384-well microtiter-plate format using either a fluorescence microplate reader at Ex/Em = 540/570 nm or a fluorescent microscope with TRITC filter.
Previously called Cellular Reactive Oxygen Species Detection Assay Kit (Orange Fluorescence).
Reactive oxygen species (ROS) are natural byproducts of the normal metabolism of oxygen and play important roles in cell signaling. However, during oxidative stress-related states, ROS levels can increase dramatically. The accumulation of ROS results in significant damage to cell structures. The role of oxidative stress in cardiovascular disease, diabetes, osteoporosis, stroke, inflammatory diseases, a number of neurodegenerative diseases and cancer has been well established. The ROS measurement will help to determine how oxidative stress modulates varied intracellular pathways.
Review the oxidative stress marker and assay guide, or the full metabolism assay guide to learn about more assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also how to assay metabolic function in live cells using your plate reader.
To measure reactive oxygen species within cells, we recommend DCFDA / H2DCFDA - Cellular ROS Assay Kit ab113851. Alternative ROS assays are available in orange (ab186028), red (ab186027), and deep red (ab186029). ab238535 is used to measure ROS in biofluids, culture supernatants and cell lysates.
For assays designed to differentiate ROS, superoxides, and reactive nitrogen species: to assay ROS and superoxides use ab139476; to assay ROS, superoxides, and reactive nitrogen species use ab139473; to assay superoxides use ab219943.
PlatformMicroplate reader, Fluor. microscope, Flow cyt.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 200 tests Assay Buffer 1 x 20ml DMSO 1 x 100µl ROS Orange Dye (Lyophilized) 1 vial
Jurkat cells were treated without (Blue) or with 100µM TBHP (Red) for 30min at 37 °C, and then loaded with ROS orange in a 5% CO2, 37 °C incubator for 1 hour. The fluorescent intensities were measured with a flow cytometer using FL2 channel
Hela cells were seeded overnight at 15,000 cells/90 µl/well in a black wall/clear bottom 96-well plate. The cells were untreated (control) or treated with1 mM H2O2 or 100 µM TBHP for 30 minutes at 37 °C. The ROS Orange assay solution (100 µl/well) was added and incubated in a 5% CO2, 37°C incubator for 1 hour. The fluorescence signal was monitored at Ex/Em = 540/570 nm (cut off = 550 nm) with bottom read mode.
Hela cells stained with the Cellular Reactive Oxygen Species Detection Assay Kit (Orange Fluorescence) (ab186028) in a black wall/clear bottom 96-well plate. A: Untreated control cells. B: Cells treated with 100 µM TBHP for 30min before staining.
ab186028 has been referenced in 3 publications.
- D'Onofrio N et al. Phenolic Profiles of Red Wine Relate to Vascular Endothelial Benefits Mediated by SIRT1 and SIRT6. Int J Mol Sci 22:N/A (2021). PubMed: 34073604
- Zhou X et al. DPP4 Inhibitor Attenuates Severe Acute Pancreatitis-Associated Intestinal Inflammation via Nrf2 Signaling. Oxid Med Cell Longev 2019:6181754 (2019). PubMed: 31827684
- Bilici K et al. Indocyanine green loaded APTMS coated SPIONs for dual phototherapy of cancer. J Photochem Photobiol B 201:111648 (2019). PubMed: 31710924